• Title/Summary/Keyword: molecular line

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NDRG2-mediated Modulation of SOCS3 and STAT3 Activity Inhibits IL-10 Production

  • Lee, Eun-Byul;Kim, Ae-Yung;Kang, Kyeong-Ah;Kim, Hye-Ree;Lim, Jong-Seok
    • IMMUNE NETWORK
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    • v.10 no.6
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    • pp.219-229
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    • 2010
  • Background: N-myc downstream regulated gene 2 (NDRG2) is a member of the NDRG gene family. Our previous report indicated a possible role for NDRG2 in regulating the cytokine, interleukin-10 (IL-10), which is an important immunosuppressive cytokine. Several pathways, including p38-MAPK, NF-${\kappa}B$, and JAK/STAT, are used for IL-10 production, and the JAK/STAT pathway can be inhibited in a negative feedback loop by the inducible protein, SOCS3. In the present study, we investigated the effect of NDRG2 gene expression on IL-10 signaling pathway that is modulated via SOCS3 and STAT3. Methods: We generated NDRG2-overexpressing U937 cell line (U937-NDRG2) and treated the cells with PMA to investigate the role of NDRG2 in IL-10 production. U937 cells were also transfected with SOCS3- or NDRG2-specific siRNAs to examine whether the knockdown of SOCS3 or NDRG2 influenced IL-10 expression. Lastly, STAT3 and SOCS3 induction was measured to identify the signaling pathway that was associated with IL-10 production. Results: RT-PCR and ELISA assays showed that IL-10 was increased in U937-mock cells upon stimulation with PMA, but IL-10 was inhibited by overexpression NDRG2. After PMA treatment, STAT3 phosphorylation was decreased in a time-dependent manner in U937-mock cells, whereas it was maintained in U937-NDRG2 cells. SOCS3 was markedly reduced in U937-NDRG2 cells compared with U937-mock cells. IL-10 production after PMA stimulation was reduced in U937 cells when SOCS3 was inhibited, but this effect was less severe when NDRG2 was inhibited. Conclusion: NDRG2 expression modulates SOCS3 and STAT3 activity, eventually leading to the inhibition of IL-10 production.

Effects of DNA Synthesis Inhibitors on the Expression of c-myc and the Stimulation of Choline Acetyltransferase Activity in Human Neuroblastoma Cell Line, IMR-32 (DNA합성 억제제가 IMR-32 세포의 c-myc 발현 및 Choline Acetyltransferase 활성도에 미치는 영향)

  • 이정은;조경혜
    • Biomedical Science Letters
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    • v.3 no.1
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    • pp.11-20
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    • 1997
  • A regulation of differentiation in human neuroblastoma cells remains poorly understood, although it is of great importance in the clinical therapy of neuroblastoma. This study was aimed to elucidate effects of DNA synthesis inhibitors on the differentiation of neuroblastoma cells on the basis of morphological, biochemical and molecular respects. Three DNA synthesis inhibitors, sodium butyrate, hydroxyurea, cytosine arabinoside were used to explore their effects on the cellular morphology, the expression of c-myc and the elevation of choline acetyltransferase activity. They led to the extension or neurite-like processes reflecting differentiation or IMR-32 cells. In addition, the treatment of three DNA synthesis inhibitors resulted in the remarkable increases in the expression of c-myc as well as the stimulation of choline acetyltransferase activity which is involved in the synthesis of acetylcholine in the differentiated cholinergic neurons. Taken together, these results indicate that DNA synthesis inhibitors play an important role in the induction of cellular differentiation in IMR-32 cells. Furthermore these DNA synthesis inhibitors seem to be future useful to give an important clue (for the treatment of neuroblastoma).

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Analysis of Studies on Ojeok-san for Establishment of Evidence Based Medicine (EBM 기반 구축을 위한 오적산 연구 논문 분석)

  • Lee, Nam-Hun;Ha, Hye-Kyung;Lee, Ho-Young;Jung, Da-Young;Choi, Ji-Yoon;Lee, Jun-Kyoung;Shin, Hyun-Kyoo
    • Herbal Formula Science
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    • v.16 no.2
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    • pp.1-9
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    • 2008
  • Objective : The purpose of this report was to provide the information about activity and safety of Ojeok-san by analyzing domestic/international papers about Ojeok-san. Methods : Domestic/international papers related to Ojeok-san were reviewed and analyzed. These papers were then classified by year, experimental method and subject. Results : The following results were obtained in this study. 1. The studies of Ojeok-san started from 1984 and has continuously increased. The studies were mainly focused on experimental models rather than clinical studies. 2. By subject, papers related to safety were most common with 5 papers among 20 papers. Besides there were papers related to efficacy of analgesic, anti-hyperlipidemic, anti-blood stasis and treatment for uterine myoma. 3. The papers related to safety were mainly focused on the effect of Okeok-san on liver function, renal function or metal concentration of organs such as blood, brain, liver, kidney and bone. Ojeok-san proved to be safe, but more clinical studies regarding the safety are needed hereafter. 4. Papers related to analgesic, anti-pyretic, anti-phlogistic activities of Ojeok-san were in vivo studies, and other papers were about anti-hyperlipidemic activity, apoptosis inducing activity on uterine myeloma cell line and anti blood static activity on hydrocortisone acetate induced blood statis model. 5. Case reports were about anti-lipidemia, analgesic effect for mastalgia/back pain and anxiety disorder due to climacteric changes. Conclusion : Ojeok-san is being used in various ways with analgesic, anti-pyretic, anti-phlogistic, anti-hyperlipidemic, anti-tumor or anti-blood statis activity. However, mechanism study should be conducted at the molecular biology level and more clinical studies on the efficacy of Ojeok-san are needed.

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Peptide Profiling and Selection of Specific-Expressed Peptides in Hypoglycemic Sorghum Seed using SELDI-TOF MS (SELDI-TOF MS를 활용한 혈당강하 수수 종자의 펩타이드 프로파일링 및 특이 발현 펩타이드 선발)

  • Park, Sei Joon;Hwang, Su Min;Park, Jun Young;Ko, Jee-Yeon;Kim, Tae Wan
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.59 no.3
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    • pp.252-262
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    • 2014
  • Sorghum seed is traditionally used as secondary food sources in addition to rice in Korea. While the hypoglycemia regulating phytochemicals have been found in sorghum seed, peptides related with hypoglycemia never been studied before. To obtain the peptide characteristics and the specifically high-expressed peptides in hypoglycemic sorghum seed, peptide profiles of seven hypoglycemic and five non-hypoglycemic sorghum lines bred in RDA were determined using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS). The twelve sorghum lines exhibited 104 peptides on CM10 protein chip array (weak cation exchange) and 95 peptides on Q10 (weak cation exchange) in the molecular mass range from 2,000 to 20,000 Da. Heat map via supervised hierarchical clustering of the significantly different peptides (p < 0.01) in peak intensity among the 12 lines effectively revealed the specifically upregulated peptides in each line and distinguished between 7 hypoglycemic and 5 non-hypoglycemic lines. Through the comparison with hypoglycemic and non-hypoglycemic lines, 10 peptides including 2231.6, 2845.4, 2907.9, 3063.5, 3132.6, 3520.8, 4078.8, 5066.2, 5296.5, 5375.5 Da were specifically high-expressed in hypoglycemic lines at p < 0.00001. This study characterized seed peptides of 12 sorghums and found ten peptides highly expressed for hypoglycemic sorghum lines, which could be used as peptide biomarkers for identification of hypoglycemic sorghum.

Application and utilization of marker assisted selection for biotic stress resistance in hybrid rice (Oryza sativa L.)

  • Song, Jae-Young;Ouk, Sothea;Nogoy, Franz Marielle;Nino, Marjohn C.;Kwon, Soon Wook;Ha, Woongoo;Kang, Kwon-Kyoo;Cho, Yong-Gu
    • Journal of Plant Biotechnology
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    • v.43 no.3
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    • pp.317-331
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    • 2016
  • Development of disease resistant plant is one of the important objectives in rice breeding programs because biotic stresses can adversely affect rice growth and yield losses. This study was conducted to identify lines with multiple-resistance genes to biotic stress among 173 hybrid rice breeding lines and germplasms using DNA-based markers. Our results showed that one hybrid rice line [IR98161-2-1-1-k1-3 (IR86409-3-1-1-1-1-1/IRBB66)] possessed 5 bacterial blight resistance genes (Xa4, xa5, Xa7, Xa13 and Xa21) while two hybrid rice lines [IR98161-2-1-1-k1-2 (IR86409-3-1-1-1-1-1/IRBB66) and 7292s (IR75589-31-27-8-33S(S1)/IR102758B)] possessed 3 bacterial blight resistance genes (Xa4, Xa7 and Xa21, and Xa3, Xa4 and xa5). Molecular survey on rice blast disease revealed that most of these lines had two different resistant genes. Only 11 lines possessed Pib, Pi-5, and Pi-ta. In addition, we further surveyed the distribution of insect resistant genes, such as Bph1, Bph18(t), and Wbph. Three hybrid breeding lines [IR98161-2-1-1-k1-3 (IR86409-3-1-1-1-1-1/IRBB66), IR98161-2-1-1-k1-2 (IR86409-3-1-1-1-1-1/IRBB66), and 7292s (IR75589-31-27-8-33S(S1) /IR102758B)] contained all three resistance genes. Finally, we obtained four hybrid rice breeding lines and germplasms [IR98161-2-1-1-k1-2 (IR86409-3-1-1-1-1-1/IRBB66), Damm-Noeub Khmau, 7290s, and 7292s (IR75589-31-27-8-33S(S1)/IR102758B)] possessing six-gene combination. They are expected to provide higher level of multiple resistance to biotic stress. This study is important for genotyping hybrid rice with resistance to diverse diseases and pests. Results obtained in this study suggest that identification of pyramided resistance genes is very important for screening hybrid rice breeding lines and germplasms accurately for disease and pest resistance. We will expand their cultivation safely through bioassays against diseases, pests, and disaster in its main export countries.

TAp73 and ΔNp73 Have Opposing Roles in 5-aza-2'-Deoxycytidine-Induced Apoptosis in Breast Cancer Cells

  • Lai, Jing;Yang, Fang;Zhang, Wenwen;Wang, Yanru;Xu, Jing;Song, Wei;Huang, Guichun;Gu, Jun;Guan, Xiaoxiang
    • Molecules and Cells
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    • v.37 no.8
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    • pp.605-612
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    • 2014
  • The p73 gene contains an extrinsic P1 promoter and an intrinsic P2 promoter, controlling the transcription of the pro-apoptotic TAp73 isoform and the anti-apoptotic ${\Delta}Np73$ isoform, respectively. The DNA methylation status of both promoters act equally in the epigenetic transcriptional regulation of their relevant isoforms. The aim of this study was to analyze the different effects of these p73 isoforms in 5-aza-2'-deoxycytidine (5-aza-dC)-induced apoptosis in breast cancer cells. We investigated the effects of the DNA demethylation agent, 5-aza-dC, on the T-47D breast cancer cell line, and evaluated the methylation status of the p73 promoters and expression of TAp73 and ${\Delta}Np73$. Furthermore, we assessed the expression of p53 and p73 isoforms in 5-aza-dC-treated T-47D cells and p53 knockout cells. 5-aza-dC induced significant anti-tumor effects in T-47D cells, including inhibition of cell viability, G1 phase arrest and apoptosis. This was associated with p73 promoter demethylation and a concomitant increase in TAp73 mRNA and protein expression. In contrast, the methylation status of promoter P2 was not associated with ${\Delta}Np73$ mRNA or protein levels. Furthermore, demethylation of P2 failed to inhibit the expression of ${\Delta}Np73$ with 5-aza-dC in the p53 knockdown cell model. Our study suggests that demethylation of the P1 and P2 promoters has opposite effects on the expression of p73 isoforms, namely up-regulation of TAp73 and down-regulation of ${\Delta}Np73$. We also demonstrate that p53 likely contributes to 5-aza-dC-induced ${\Delta}Np73$ transcriptional inactivation in breast cancer cells.

Suppression of Thrombospondin-1 Expression by PMA in the Porcine Aortic Endothelial Cells (정상 돼지 대동맥 내피세포에서 PMA에 의한 thrombospondin-1 발현 억제)

  • Chang, Seo-Yoon;Kang, Jung-Hoon;Hong, Kyong-Ja
    • Journal of Life Science
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    • v.14 no.1
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    • pp.154-162
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    • 2004
  • Thrombospondin-1 (TSP-1), a negative regulator in tumor growth and angiogenesis, is cell-type specifically regulated and at transcriptional level by external stimuli. Previously, we found that phorbol 12-myristate 13-acetate (PMA) suppressed TSP-1 expression in porcine aortic endothelial (PAE) cell, but enhanced in hepatoma cell line, Hep 3B cell. A region between -767 and -723 on the tsp-1 promoter was defined as a responsive site to the suppression in PAE cell. eased on the previous results, the molecular mechanism of TSP-1 expression was determined by characterizing interactions between cis-elements and trans-factors using three overlapped oligonucleotide probes, oligo a-1 (from -767 to -738), a-2 (-759 to -730) and a-3 (-752 to -723). The results from electromobility shift assay showed that PMA-induced suppression of TSP-1 transcription in PAE cell might be caused via a negative regulator binding to the region from -752 to -730 and additionally generated by lacking two positive regulators binding to the sites from -767 to -760 and from -752 to -730. Especially, PMA enhanced the binding ability of the negative regulator to the site from -752 to -730 in PAE cell, but anti-c-Jun did not affected its binding ability.

Preliminary Study on the Enhancement of Reconstruction Speed for Emission Computed Tomography Using Parallel Processing (병렬 연산을 이용한 방출 단층 영상의 재구성 속도향상 기초연구)

  • Park, Min-Jae;Lee, Jae-Sung;Kim, Soo-Mee;Kang, Ji-Yeon;Lee, Dong-Soo;Park, Kwang-Suk
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.5
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    • pp.443-450
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    • 2009
  • Purpose: Conventional image reconstruction uses simplified physical models of projection. However, real physics, for example 3D reconstruction, takes too long time to process all the data in clinic and is unable in a common reconstruction machine because of the large memory for complex physical models. We suggest the realistic distributed memory model of fast-reconstruction using parallel processing on personal computers to enable large-scale technologies. Materials and Methods: The preliminary tests for the possibility on virtual manchines and various performance test on commercial super computer, Tachyon were performed. Expectation maximization algorithm with common 2D projection and realistic 3D line of response were tested. Since the process time was getting slower (max 6 times) after a certain iteration, optimization for compiler was performed to maximize the efficiency of parallelization. Results: Parallel processing of a program on multiple computers was available on Linux with MPICH and NFS. We verified that differences between parallel processed image and single processed image at the same iterations were under the significant digits of floating point number, about 6 bit. Double processors showed good efficiency (1.96 times) of parallel computing. Delay phenomenon was solved by vectorization method using SSE. Conclusion: Through the study, realistic parallel computing system in clinic was established to be able to reconstruct by plenty of memory using the realistic physical models which was impossible to simplify.

Inhibition of Production of Reactive Oxygen Species and Gene Expression Profiles by Cirsii Japonici Herba Extract Treatment in HepG2 Cells

  • Rho, Sam-Woong;Chung, Hwan-Suck;Kang, Moon-Kyu;Na, Young-In;Cho, Chong-Woon;Kim, Hyung-Min;Jung, Hyuk-Sang;Park, Hi-Joon;Kim, Hong-Yeoul;Hong, Moo-Chang;Shin, Min-Kyu;Kim, Sung-Soo;Bae, Hyun-Su
    • Molecular & Cellular Toxicology
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    • v.1 no.4
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    • pp.224-229
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    • 2005
  • Cirsii Japonici Herba (CJH) extract has been used for hundreds of years in Asian countries as a treatment for pollutant, radiation, and alcohol-induced liver damage. The reducing effect of CJH on hydrogen peroxide-induced reactive oxygen species (ROS) production, the main cause of cell damage or death, was evaluated using the HepG2 cell line. Cell survival was determined using MTS assay. The viability of cells treated with CJH was not significantly different from oxidative-stressed HepG2 cells. A dose-dependent inhibitory effect by CJH on ROS production was shown in oxidative-stressed cells using the $H_{2}DCFDA$ assay. To identify candidate genes responsible for the anti-oxidative effects of CJH on HepG2 cells, an oligonucleotide microarray analysis was performed. The expressions of five genes were decreased, whereas nineteen genes were up-regulated in CJH plus hydrogen peroxide treated cells, compared to only hydrogen peroxide treated cells. Among them, the expression of 5 genes was decreased in hydrogen peroxide treatment when compared to control. These genes are known to regulate cell survival and progression. On the other hand, it was shown that its main compounds were not a sylimarin or its analogs. The list of differentially expressed genes may provide further insight on the action and mechanism behind the anti-oxidative effects of Cirsii Japonici Herba.

Characterization of a Fibrinolytic Enzyme from Pickled Anchovy (멸치젓갈유래의 혈전용해호소에 대한 특성)

  • Yang Woong-Suk;Lim Hak-Seob;Chung Kyung Tae;Kim Young-Hee;Huh Man Kyu;Choi Byung Tae;Choi Yung Hyun;Jeong Yong Kee
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.434-438
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    • 2005
  • In the previous study, we isolated a myulchikinase (MK), which has fibrinolytic activity and cytotoxicity to the tumor cell line, from myl- chi-jeot-gal. In this study, the effect of NaCl concentration, metallic ions, pH, temperature, and plasminogen on the activity of MK was analysed. The MK activity was maintained at least $80\%$ activity up to $30\%$ NaCl, which indicates that the enzyme may be halotolerant. The optimal pH and temperature were 8 and $40^{\circ}C$, respectively. The fibrinolytic activity of MK was completely inhibited with 0.5 mM $Hg^{2+}$ and inhibited to $50^{\circ}C$ with 1 mM $Cu^{2+}\;and\;Zn^{2+}$. The MK showed strong activity in plasminogen- rich fibrin plate but not in plasminogen-free fibrin plate. The result indicates that the MK may be a plasminogen activator type fibrinolytic enzyme.